猪DFAT细胞成脂再分化过程中KLF2和PPARγ的表达

发布时间：2018-10-16 15:45

【摘要】：旨在探讨猪去分化脂肪细胞(DFAT)再分化过程中Krüppel样因子2(KLF2)和过氧化物酶体增殖物激活受体γ(PPARγ)的表达模式,为揭示猪DFAT细胞再分化机制奠定基础。采集猪成熟脂肪细胞,经"天花板"培养法获得DFAT细胞,用流式细胞仪检测表面抗原,形态学和油红O染色提取法检测成脂分化程度,Real-time PCR检测KLF2和PPARγmRNA的表达。结果表明,猪成熟脂肪细胞接种第3天,可见明显的去分化迹象,即细胞形成犄角状突起,大脂滴分解成小脂滴并向细胞外排出脂滴,接种至第9天脂滴基本排出完毕,接种至第12天原代细胞长满底壁;间充质干细胞表面抗原CD29、CD44和CD105的阳性表达率分别达到99.0%、97.8%和99.5%,而造血干细胞表面抗原CD45和CD34的表达仅为3.55%和4.55%;将F3代细胞成脂诱导3d,胞质中出现脂滴,并随诱导时间增加,脂滴数量增多,体积增大,诱导至12d诱导率达80%以上;成脂诱导2、5、10和15d时,KLF2mRNA的相对表达量分别为0.47±0.02、0.35±0.07、0.31±0.09和0.11±0.07,PPARγmRNA的相对表达量分别为1.62±0.01、2.03±0.04、3.22±0.04和3.49±0.06。本研究成功获得高纯度的DFAT细胞,具有间充质干细胞特性和成脂再分化能力;KLF2 mRNA的表达在成脂分化过程中随诱导时间的增加而减少,而PPARγmRNA的表达量随诱导时间的延长逐步增加;表明KLF2在DFAT细胞的成脂再分化过程中可能发挥抑制作用,而PPARγ可能会发挥促进作用。
[Abstract]:The aim of this study was to investigate the expression patterns of Peppel-like factor 2 (KLF2) and peroxidase proliferator-activated receptor (CDR3) in the redifferentiation of porcine dedifferentiated adipocytes (DFAT), and lay a foundation for revealing the differentiation mechanism of DFAT cells in pigs. Porcine mature adipocytes were collected, DFAT cells were obtained by "Ceiling" culture, surface antigen, morphology and oil-red O staining were detected by flow cytometry to detect the degree of lipid differentiation. Real-time PCR was used to detect the expression of KLF2 and Jurkat mRNA. The results showed that the mature fat cells of pigs were inoculated on day 3, and there were obvious signs of dedifferentiation, that is, the cells were formed into small angle-shaped protrusions, the big fat drops were decomposed into small fat drops and the fat drops were discharged out of the cells, and the lipid droplets inoculated to day 9 were basically discharged, and the primary cells inoculated to the 12th day were overgrown with the bottom wall. The positive expression rates of CD29, CD44 and CD105 were 99. 0%, 97. 8% and 99. 5%, respectively, while the expression of CD45 and CD34 in hematopoietic stem cells was only 3.55% and 4.55%. The relative expressions of KLF2mRNA were 0. 47, 0. 02, 0. 35, 0. 07, 0. 31, 0. 09 and 0. 11, 0. 07, 0. 31, 0. 09 and 0. 11, 0. 07, 0. 31, 0. 09 and 0. 11, 0. 07, 0. 31, 0. 09 and 0. 11, 0. 07, 3. 22, 0. 04 and 3. 49, 0. 06, respectively. The results showed that the expression of KLF2 mRNA decreased with the increase of induction time in the process of lipodifferentiation, while the expression of KLF2 mRNA increased with the increase of induction time. It was suggested that KLF2 might play an inhibitory role in the process of fat redifferentiation of DFAT cells, and the effect of KLF2 could play a role in promoting the differentiation of FAT cells.
【作者单位】：青岛农业大学动物科技学院;
【基金】：国家自然科学基金项目(31372392)
【分类号】：S828

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